Leiomyosarcoma of the Urinary Bladder in Adult Patients: A Systematic Review of the Literature and Meta-Analysis.
Leiomyosarcoma of the urinary bladder is exceedingly rare. Most clinicians come across only a few cases during their career, and information regarding treatment and outcome is scattered in the scientific literature. Interested clinicians and patients have to undertake troublesome search for treatment and outcome information. We performed a systematic review of the literature using the PubMed and Web of Science databases and included all identified cases published in English language between 1970 and June 2018 into a meta-analysis. Prior to the literature search, key questions were formulated and with the data obtained, answers to these questions should be derived. We analyzed clinical data of 210 cases of urinary bladder leiomyosarcoma revealed by this review and seen in our institution. The mean age of patients was 52 years. The majority (75%) of the tumors was classified as high-grade sarcomas. We found no report of a prior radiation therapy to the pelvic organs, but some authors suggested an association between cyclophosphamide treatment and the development of bladder leiomyosarcoma, especially in patients with retinoblastoma. For the whole sample, we determined 5- and 10-year cancer-specific cumulative mortality rates of 38 and 50%. Patients with high-grade sarcomas had a trend toward a higher mortality compared with low-grade tumors (p = 0.0280). The most promising treatment option seems to be surgery (radical or partial cystectomy) with negative resection margins, possibly supplemented by chemotherapy or radiation. About half of patients with bladder leiomyosarcoma survived on the long run. Low-grade tumors may have a better outcome with, nevertheless, countable long-term mortality. For better assessment of that rare bladder tumor, its best treatment options, and the influence of neoadjuvant or adjuvant therapies on the outcome of patients, a larger series with long-term survival data is required.


Fabrication of hydrophobic NiFe2O4@poly(DVB-LMA) sponge via a Pickering emulsion template method for oil/water separation.
Super-hydrophobic porous absorbents are convenient, low-cost, efficient and environment-friendly materials in the treatment of oil spills. In this work, a simple Pickering emulsion template method was employed to fabricate an interconnected porous poly(DVB-LMA) sponge. A new co-Pickering stabilization system of Span 80 and NiFe2O4 nanoparticles was used to prepare ultra-concentrated internal phase water-in-oil (W/O) emulsions. After further polymerization, the resulting sponges were generated, which exhibited excellent adsorption selectivity due to the super-hydrophobicity and super-lipophilicity. Furthermore, the characterization results indicated that the composites had superior thermal stability, low density, high porosity and a flexible three-dimensional porous structure. Besides, the addition of nickel ferrite nanoparticles provided the materials with extra magnetic operability. High oil adsorption capacity (up to 36.9-84.2 g g-1), high oil retention, fast adsorption rate and superior reusability allowed the materials to be applied in the treatment of oily water.


Mode discrimination in dissipatively coupled laser arrays.
We show that dissipative coupling between an array of passive optical resonators creates a ladder of decay rates in the complex eigenfrequencies. This effect promotes mode discrimination in laser arrays, while the lowest-order and highest-order modes exhibit the highest and lowest lasing thresholds, respectively. The array supermodes and their corresponding eigenfrequencies are calculated analytically through a tight-binding model, and the single-mode operation range is derived. The results are exemplified through the finite element simulation of an array of transversely coupled semiconductor laser cavities.


Hypoxia associated biomarkers in lung cancer - an update.
The high incidence rates of lung cancer have resulted in multiple progressions in cancer research leading to the development of novel therapeutic strategies. The three main branches of cancer research where improvements are being made are surgery, chemotherapy, and radiotherapy. These research developments have significantly improved the survival rates of cancer patients as most of these therapeutic options target specific molecules involved in cancer progression and metastasis. Experimental research has successfully identified potentially important molecules responsible for cancer progression. Further, the above molecular markers of disease are clinically approved biomolecules of significant prognostic value. The above molecular biomolecules also helped in rapid identification of subgroups of patients who need immediate emergency treatment. Hypoxia is one of such conditions where the human system is deprived of an adequate oxygen supply and requires immediate treatment. Hypoxia markers have been proved to be beneficial in the timely management of cancer. The present review is focused on the latest updates in the area of hypoxia related to markers in the prognosis of lung cancer.


A screening programme for hypertension in general practice.
It is recommended that hypertensive patients with a diastolic blood pressure of 100 mmHg be treated. However, it has been proposed that only 50% of hypertensives are diagnosed, and of these, 50% are not treated. We therefore set out to identify hypertensive patients in our practice, who were then actively treated, some in a clinical trial of ketanserin. Of the 3,384 patients (1,667 males and 2,707 females) who were invited to our surgery for a blood pressure measurement, 2,606 patients (77%) attended. The overall prevalence of hypertension in the patients, aged 35-65 years was 2.9%. This prevalence increased with age: from 1.0% in the 35-44 year age group to 3.5% at 45-54 years, and 4.4% at 55-64 years. The mean systolic but not diastolic blood pressure increased with age. There were more male hypertensives than females, except over 55 years of age. Of the 84 hypertensives identified after one visit, the majority remained hypertensive after an additional visit, and 31 agreed to participate in a clinical trial. After a four-week placebo run-in, 22 patients had a diastolic blood pressure above 95 mmHg and were randomly allocated to receive ketanserin 40 mg twice daily or metoprolol 100 mg twice daily. Treatment continued double-blind for three months. There were no significant differences in blood pressure reduction between the treatment groups. The heart rate was reduced more by metoprolol. There were no withdrawals for side effects and no major differences in subjective complaints between the two treatment groups.


Genetic diversity in the Helicobacter pylori cag pathogenicity island and effect on expression of anti-CagA serum antibody in UK patients with dyspepsia.
To investigate variation within the cag pathogenicity island (PAI) of Helicobacter pylori isolated from patients with dyspepsia in mid-Essex, and to evaluate the effect on expression of anti-CagA antibody. Sixty two isolates of H pylori cultured from gastric biopsies were screened by specific PCR assays for the presence of cagA and other gene markers (cagD and cagE, and virD4) in the cag PAI. An enzyme linked immunosorbent assay (ELISA) kit (Viva Diagnostica helicobacter p120) was used to test for anti-CagA IgG antibody in matching sera. Isolates were also genotyped by vacuolating cytotoxin polymerase chain reaction (PCR) analysis, and tested for absence of the complete cag PAI (empty site PCR assay). Forty one of the H pylori isolates had a cag PAI containing cagA. One strain had no cagA but other cag PAI loci were present, whereas the remaining 20 strains had no detectable cag PAI markers. Anti-CagA IgG antibody was detected in 34 sera by the ELISA assay, and when compared with the cag PAI genotype of the infecting strain, accuracy, sensitivity, and specificity were 92%, 87%, and 100%, respectively. The seven discrepant or borderline strains in the ELISA were all vacA s1 but differed in other genotypic markers. The cag PAI was widely distributed in H pylori from patients with dyspepsia in mid-Essex who had different gastric pathologies. Infection with a strain having an uninterrupted cag PAI was associated with the presence of anti-CagA antibody in most patients. Discrepant ELISA results, mostly for elderly patients with duodenal ulcers, were attributed to cagA associated variation, particularly to the presence of mixed cagA+/cagA- cell variants in the infecting strain population. Tests for anti-CagA serum antibody were unreliable for predicting severity of clinical disease associated with H pylori infection in this series of patients.


Hypersensitivity in the anterior median eye of a jumping spider.
Changes in sensitivity of the photoreceptor cells of the anterior median eye of the jumping spider Menemerus confusus Boes. et Str. have been studied by recording electroretinograms (ERGs) and receptor potentials. The amplitudes of the responses (ERGs and receptor potentials) increase during repetitive stimulation, with a maximum increase at 3-5 s intervals. The sensitivity of the photoreceptor cell is greater for about 60 s following illumination (maximum magnitude at 3-5 s) than it is during complete dark adaptation. This phenomenon, which we call 'hypersensitivity', is lost within one day following surgery in physiological saline. Upon loss of hypersensitivity, the sensitivity decrease during light adaptation is greater than for the normal eye and the small increase of sensitivity following the onset of illumination observed for the normal eye is lost.


Surgical treatment of unstable fractures of the distal clavicle: a comparative study of Kirschner wire and clavicular hook plate fixation.
During 1988-1999 39 unstable fractures of the distal clavicle (Neer 2) were operated on in Oulu University Hospital. Kirschner wire (K-wire) fixation was used in 22 cases and a clavicular hook plate in 17. Shoulder symptoms and function were assessed using self-administered questionnaires devised by L'Insalata et al. (1997), and Constant scoring. Mean follow-up was 6.2 years in the K-wire fixation group and 2.0 years in the clavicular hook plate one. The mean L'Insalata scores were 91 in both groups (92% and 93% of the contralateral side) and the mean Constant scores 84 (95% and 90 (96%) for K-wire fixation and the clavicular hook plate, respectively. Complications commonly occurred with K-wires, which migrated in 12 cases, resulting in loss of reduction in 7 and infection in 3, and 2 cases of non-union. In the clavicular hook plate group, there was 1 complication, a fracture of the clavicle, and 2 cases of non-union. We conclude that shoulder symptoms were reduced and function restored to an adequate level by both methods, but complications were unacceptably frequent when K-wires were used. The clavicular hook plate was better in this respect and it is therefore recommended.


Nuclear pre-mRNA decapping and 5' degradation in yeast require the Lsm2-8p complex.
Previous analyses have identified related cytoplasmic Lsm1-7p and nuclear Lsm2-8p complexes. Here we report that mature heat shock and MET mRNAs that are trapped in the nucleus due to a block in mRNA export were strongly stabilized in strains lacking Lsm6p or the nucleus-specific Lsm8p protein but not by the absence of the cytoplasmic Lsm1p. These nucleus-restricted mRNAs remain polyadenylated until their degradation, indicating that nuclear mRNA degradation does not involve the incremental deadenylation that is a key feature of cytoplasmic turnover. Lsm8p can be UV cross-linked to nuclear poly(A)(+) RNA, indicating that an Lsm2-8p complex interacts directly with nucleus-restricted mRNA. Analysis of pre-mRNAs that contain intronic snoRNAs indicates that their 5' degradation is specifically inhibited in strains lacking any of the Lsm2-8p proteins but Lsm1p. Nucleus-restricted mRNAs and pre-mRNA degradation intermediates that accumulate in lsm mutants remain 5' capped. We conclude that the Lsm2-8p complex normally targets nuclear RNA substrates for decapping.


Is viral E6 oncoprotein a viable target? A critical analysis in the context of cervical cancer.
An understanding of the pathology of cervical cancer (CC) mediated by E6/E7 oncoproteins of high-risk human papillomavirus (HPV) was developed by late 80's. But if we look at the present scenario, not a single drug could be developed to inhibit these oncoproteins and in turn, be used specifically for the treatment of CC. The readers are advised not to presume the "viability of E6 protein" as mentioned in the title relates to just druggability of E6. The viability aspect will cover almost everything a researcher should know to develop E6 inhibitors until the preclinical stage. Herein, we have analysed the achievements and shortcomings of the scientific community in the last four decades in targeting HPV E6 against CC. Role of all HPV proteins has been briefly described for better perspective with a little detailed discussion of the role of E6. We have reviewed the articles from 1985 onward, reporting in vitro inhibition of E6. Recently, many computational studies have reported potent E6 inhibitors and these have also been reviewed. Subsequently, a critical analysis has been reported to cover the in vitro assay protocols and in vivo models to develop E6 inhibitors. A paragraph has been devoted to the role of public policy to fight CC employing vaccines and whether the vaccine against HPV has quenched the zeal to develop drugs against it. The review concludes with the challenges and the way forward.


Neurovascular Unit: A New Target for Treating Early Stages of Diabetic Retinopathy.
The concept of diabetic retinopathy as a microvascular disease has evolved and is now considered a more complex diabetic complication in which neurovascular unit impairment plays an essential role and, therefore, can be considered as a main therapeutic target in the early stages of the disease. However, neurodegeneration is not always the apparent primary event in the natural story of diabetic retinopathy, and a phenotyping characterization is recommendable to identify those patients in whom neuroprotective treatment might be of benefit. In recent years, a myriad of treatments based on neuroprotection have been tested in experimental models, but more interestingly, there are drugs with a dual activity (neuroprotective and vasculotropic). In this review, the recent evidence concerning the therapeutic approaches targeting neurovascular unit impairment will be presented, along with a critical review of the scientific gaps and problems which remain to be overcome before our knowledge can be transferred to clinical practice.


Tissue Classification During Needle Insertion Using Self-Supervised Contrastive Learning and Optical Coherence Tomography.
Needle positioning is essential for various medical applications such as epidural anaesthesia. Physicians rely on their instincts while navigating the needle in epidural spaces. Thereby, identifying the tissue structures may be helpful to the physician as they can provide additional feedback in the needle insertion process. To this end, we propose a deep neural network that classifies the tissues from the phase and intensity data of complex OCT signals acquired at the needle tip. We investigate the performance of the deep neural network in a limited labelled dataset scenario and propose a novel contrastive pretraining strategy that learns invariant representation for phase and intensity data. We show that with 10% of the training set, our proposed pretraining strategy helps the model achieve an F1 score of 0.84±0.10 whereas the model achieves an F1 score of 0.60±0.07 without it. Further, we analyse the importance of phase and intensity individually towards tissue classification.


Tuberous sclerosis complex. Forty-years of follow-up of a patient affected.
To report the case of a patient diagnosed with tuberous sclerosis complex (TSC), describe its clinical features, diagnosis, and to attract attention on the fact that after 40 years of follow-up, the patient has presented practically all the manifestations described in the literature. A 42-year-old man diagnosed with.TSC presented the emergency department due to left lumbar pain and self-limited gross hematuria. On clinical examination patient was haemodynamically stable, but with decrease in haemoglobin (6.8 g/dL). Abdominal CT scan showed a 20 cm diameter heterogeneous mass in the left kidney suggesting hemorrhage of an angiomyolipoma. Left radical nephrectomy was performed and the pathological study of the surgical specimen confirmed the diagnosis of angiomyolipoma. Inmunohistochemical staining was positive with HMB-45. To recommend that patients with TSC be evaluated by a multidisciplinary group of clinicians, including urologists, neurologists and dermatologists. As patients with TSC survive into adulthood they will require more intervention by the urologist. CT scan is usually enough for the diagnosis of angiomyolipomas. Complete nephrectomy is appropriate when the whole kidney has been replaced by angiomyolipoma. The identification of molecular markers (HMB-45) facilitates histopathological diagnosis.


[Antagonism of baicalin on cell cyclical distribution and cell apoptosis in A549 cells infected with influenza A (H1N1) virus].
To observe the inhibitive effect of Baicalin against influenza A H1N1 virus infection in epithelial cell line A549, the cell proliferation and cytotoxicity were assayed by MTT, the cell cycle and the apoptosis were analyzed by flowcytometer using PI staining, the morphology of cellular nucleolus was observed by Hoechst 33258 staining and the effects of activation on caspase 3 and caspase 8/9 were also detected by immunofluorescent staining with a fluorescence microscope. The results showed that Baicalin exerted an inhibitive effect on CPE after influenza A H1N1 virus infection. The FACS with PI staining showed that the cell cycle of the infected cell was arrested at S phase, the Baicalin-treated group decreased S phase cell ratio and subG0 phase peak in comparison with the control (P < 0.05) and significantly promoted cell proliferation (# P < 0.05). Hoechst33258 staining suggested that Baicalin protected the cellular nucleolus against the influenza virus-induced apoptosis. Observation under the immunofluorescent microscope suggested that the activities of caspase-8 and caspase-3 were enhanced at 36 h post the influenza virus infection, but 100 microg/mL Baicalin suppressing the activation of caspase-8 and caspase-3 rather than that of caspase-9. In summary, this research confirmed that Baicalin inhibited the influenza A H1N1 virus strain infection in vitro, the drug obviously protected cells from apoptosis damages through regulating cell cycle and suppressed the activation of caspase-8 and caspase-3. The down-regulation was significant and showed a dose-dependent relationship.


[Acute prostatitis as the 1st symptom of brucellosis].
To report a case of acute prostatitis as the first symptom of brucellosis. 43-year-old patient who presented with a clinical picture of acute prostatitis and a febrile syndrome for seven days. Physical exam, blood and urine analysis and microbiological tests, and specific serologic studies were performed. Microbiological diagnosis was negative for bacteria, but Bengala pink (+), antibrucella serum agglutination (+) 1/320, and anti-Brucella Coombs test (+) 1/5120. Specific treatment was started with doxycyclin 100 mg b.i.d for 45 days and streptomycin 1 gr o.d for 15 days, with a positive outcome. The genitourinary tract is the second most frequent localization, being the testicle the most frequently affected organ, although it can also be localized in other areas of the urinary tract.


Papillary thyroid carcinoma involving cervical neck lymph nodes: correlations with lymphangiogenesis and ultrasound features.
Stratification of risk factors for cervical lymph node metastasis (LNM) in thyroid papillary carcinoma is important for providing standards for post-operative adjuvant radio-iodine therapy and for patient prognosis. We investigated pathological factors based on the lymphatic vessel system and radiological features associated with tumor with cervical neck LNM. Among patients who had undergone thyroidectomy confirmed to be papillary thyroid carcinoma, we selected 126 age-sex matched paired patients without cervical LNM (group 1) and with LNM (group 2) to evaluate risk factors. Pathological factors evaluated were size, multiplicity, and extra thyroid extension state, based on the pathological reports using stored data. The lymphatic vessel density (LVD) of each tumor was evaluated by staining for VEGFR-3 and D2-40 and correlated with cervical LNM state. Malignant ultrasound features were evaluated to compare the differences between these two groups. Larger tumor size, multiplicity, extrathyroid extension were more common in group 2 (p<0.05). The median percentage of VEGFR-3 for group 1 was 20 (range 0-30) and D2-40 was 13 (range 7-23) while for group 2, VEGFR-3 was 80 (70-90) and D2-40 was 78 (54-114). LVD measured by intratumoral D2-40 staining was 20.6% and 79.4% for group 1 and group 2, respectively. Intra-tumoral lymphatics measured by D2-40 stain had a strong correlation with cervical LNM (Odds 1.230, CI 1.01.-1.499 p value 0.040). Ultrasound (US) features had no significant differences between the two groups although calcifications tended to be higher in group 2 (84% vs. 76% p=0.264). Lymphatic vessel density and nodule echogenicity were not associated with LNM. Intratumoral lymphangiogenesis was most strongly associated with LNM and thus, could be a useful predictive marker for cervical LNM.


A systematic assessment of current genome-scale metabolic reconstruction tools.
Several genome-scale metabolic reconstruction software platforms have been developed and are being continuously updated. These tools have been widely applied to reconstruct metabolic models for hundreds of microorganisms ranging from important human pathogens to species of industrial relevance. However, these platforms, as yet, have not been systematically evaluated with respect to software quality, best potential uses and intrinsic capacity to generate high-quality, genome-scale metabolic models. It is therefore unclear for potential users which tool best fits the purpose of their research. In this work, we perform a systematic assessment of current genome-scale reconstruction software platforms. To meet our goal, we first define a list of features for assessing software quality related to genome-scale reconstruction. Subsequently, we use the feature list to evaluate the performance of each tool. To assess the similarity of the draft reconstructions to high-quality models, we compare each tool's output networks with that of the high-quality, manually curated, models of Lactobacillus plantarum and Bordetella pertussis, representatives of gram-positive and gram-negative bacteria, respectively. We additionally compare draft reconstructions with a model of Pseudomonas putida to further confirm our findings. We show that none of the tools outperforms the others in all the defined features. Model builders should carefully choose a tool (or combinations of tools) depending on the intended use of the metabolic model. They can use this benchmark study as a guide to select the best tool for their research. Finally, developers can also benefit from this evaluation by getting feedback to improve their software.


Clinical effects of myotonic dystrophy on pregnancy and the neonate.
The article describes pregnancies and labors of five women with myotonic muscular dystrophy and their four severely involved infants, and reviews the pertinent literature. Three of the four neonatal cases died by 3 weeks of age of respiratory failure or aspiration; the fourth infant is now 4 months old and has respiratory and swallowing difficulties. The symptoms of myotonic dystrophy worsen during pregnancy. A high rate of fetal loss occurs due to spontaneous abortion, prematurity, and neonatal involvement with the disease. Prolonged labor has been described as a consistent complication, but the evidence does not justify this conclusion. Although many neonates with myotonic dystrophy are asymptomatic, severely affected newborns have a recognizable disorder unrelated to the severity of the maternal disease. The most common clinical manifestations in the neonate are arthrogryposis involving predominantly the lower extremilies, generalized hypotonia and weakness, and pharyngeal weakness. Less constant features include polyhydramnios, facial diplegia, diaphragmatic paralysis, respiratory failure, decreased motility of the gastrointestinal tract, congenital cataracts, and electrocardiographic abnormalities.


Role, Laboratory Assessment and Clinical Relevance of Fibrin, Factor XIII and Endogenous Fibrinolysis in Arterial and Venous Thrombosis.
Diseases such as myocardial infarction, ischaemic stroke, peripheral vascular disease and venous thromboembolism are major contributors to morbidity and mortality. Procoagulant, anticoagulant and fibrinolytic pathways are finely regulated in healthy individuals and dysregulated procoagulant, anticoagulant and fibrinolytic pathways lead to arterial and venous thrombosis. In this review article, we discuss the (patho)physiological role and laboratory assessment of fibrin, factor XIII and endogenous fibrinolysis, which are key players in the terminal phase of the coagulation cascade and fibrinolysis. Finally, we present the most up-to-date evidence for their involvement in various disease states and assessment of cardiovascular risk.


A randomised controlled trial of the Buteyko technique as an adjunct to conventional management of asthma.
To assess the effectiveness of a non-pharmacological intervention in patients with asthma on conventional therapy including inhaled corticosteroid. A randomised controlled trial of the Buteyko technique in a group of adults with asthma. The control group was trained by a physiotherapist in breathing and relaxation techniques. A single centre associated with a University-based asthma programme. Asthma control, defined by a composite score based on the Canadian asthma consensus report 6 months after completion of the intervention. Both groups showed substantial and similar improvement and a high proportion with asthma control 6 months after completion of the intervention. In the Buteyko group the proportion with asthma control increased from 40% to 79% and in the control group from 44% to 72%. In addition the Buteyko group had significantly reduced their inhaled corticosteroid therapy compared with the control group (p=0.02). None of the other differences between the groups at 6 months were significant. Six months after completion of the interventions, a large majority of subjects in each group displayed control of their asthma with the additional benefit of reduction in inhaled corticosteroid use in the Buteyko group. The Buteyko technique, an established and widely recognised intervention, or an intensive programme delivered by a chest physiotherapist appear to provide additional benefit for adult patients with asthma who are being treated with inhaled corticosteroid.


Continuous phase regulation of MoSe2 from 2H to 1T for the optimization of peroxidase-like catalysis.
Nanozymes are a new generation of artificial enzymes that address the limitations of natural enzymes, with numerous reports on the development of high performance nanozymes for various applications. Herein, the phase regulation of network-like MoSe2 from the metal 1T phase to semiconductor 2H phase was achieved under different reaction conditions by a facile but efficient microwave-assisted solvothermal method. The coexisting 1T/2H-MoSe2 with edge enriched and defective nanostructures showed great enhancement of the peroxidase-like properties. This high enzymatic activity may arise from optimized active sites of 1T/2H-MoSe2, effectively leading to a rapid charge transfer within the substrate. In addition, chitosan functionalization not only greatly improves the dispersion stability of MoSe2, but also significantly increases its peroxidase-like activity, probably due to its enhanced affinity to hydrogen peroxide (H2O2). Based on these results we have established a highly sensitive colorimetric assay for the detection of H2O2 and glucose in human serum.


Comparison of the iLUX and the LipiFlow for the Treatment of Meibomian Gland Dysfunction and Symptoms: A Randomized Clinical Trial.
To compare the effects of eyelid treatment with the iLUX MGD Treatment System and the LipiFlow Thermal Pulsation System on objective and subjective parameters of meibomian gland function and symptoms. In this randomized, open-label, controlled, multicenter clinical trial, both eyes of 142 patients aged ≥18 years with Ocular Surface Disease Index (OSDI) scores ≥23, total meibomian gland scores (MGS) ≤12 in the lower eyelid of each eye, and tear break-up time (TBUT) <10 s were randomized 1:1 to iLUX or LipiFlow treatment, with stratification by test center. The primary effectiveness endpoints were changes in total MGS (masked) and TBUT from baseline to 4 weeks. The secondary effectiveness endpoint was changed in OSDI score from baseline to 4 weeks. Both devices significantly improved effectiveness outcomes, with no differences between the two devices. At the 4-week visit, mean MGS, TBUT, and OSDI scores improved at least 16.9 ± 11.5, 2.6 ± 3.2 s, and 28.0 ± 22.8, respectively, across treatment groups and treated eyes. Four device/procedure-related events occurred in the iLUX group, compared with none in the LipiFlow group, but there were no device-related adverse events that involved changes in lid margins, eyelids, or lash integrity. Corneal staining, intraocular pressure, and visual acuity did not differ in the two groups. Both treatments produced significant improvements in meibomian gland function and symptoms. For all effectiveness measures, there were no statistically significant differences between the two treatments.


Single-molecule approaches to prion protein misfolding.
The structural conversion of the prion protein PrP into a transmissible, misfolded form is the central element of prion disease, yet there is little consensus as to how it occurs. Key aspects of conversion into the diseased state remain unsettled, from details about the earliest stages of misfolding such as the involvement of partially- or fully-unfolded intermediates to the structure of the infectious state. Part of the difficulty in understanding the structural conversion arises from the complexity of the underlying energy landscapes. Single molecule methods provide a powerful tool for probing complex folding pathways as in prion misfolding, because they allow rare and transient events to be observed directly. We discuss recent work applying single-molecule probes to study misfolding in prion proteins, and what it has revealed about the folding dynamics of PrP that may underlie its unique behavior. We also discuss single-molecule studies probing the interactions that stabilize non-native structures within aggregates, pointing the way to future work that may help identify the microscopic events triggering pathogenic conversion. Although single-molecule approaches to misfolding are relatively young, they have a promising future in prion science.


Effect of Partial Crystallization on the Structural and Luminescence Properties of Er3+-Doped Phosphate Glasses.
Er-doped phosphate glass ceramics were fabricated by melt-quenching technique followed by a heat treatment. The effect of the crystallization on the structural and luminescence properties of phosphate glasses containing Al₂O₃, TiO₂, and ZnO was investigated. The morphological and structural properties of the glass ceramics were characterized by Field Emission-Scanning Electron Microscopy (FE-SEM), X-ray Diffraction (XRD), and micro-Raman spectroscopy. Additionally, the luminescence spectra and the lifetime values were measured in order to study the influence of the crystallization on the spectroscopic properties of the glasses. The volume ratio between the crystal and the glassy phases increased along with the duration of the heat treatment. The crystallization of the glass ceramics was confirmed by the presence of sharp peaks in the XRD patterns and different crystal phases were identified depending on the glass composition. Sr(PO₃)₂ crystals were found to precipitate in all the investigated glasses. As evidenced by the spectroscopic properties, the site of the Er3+ ions was not strongly affected by the heat treatment except for the fully crystallized glass ceramic which does not contain Al₂O₃, TiO₂, and ZnO. An increase of the lifetime was also observed after the heat treatment of this glass. Therefore, we suspect that the Er3+ ions are incorporated in the precipitated crystals only in this glass ceramic.


Severe Neck Pain with Fever: Is it Meningitis?
A 58-year-old male patient presented to the emergency department with complaints of severe neck pain. He admitted to drug use but denied using intravenous (IV) drugs. On exam, he had a fever of 100.7°F, positive Kernig's sign, and normal neurologic exam. The patient was suspected to have bacterial meningitis and was started on IV antibiotics. The next day the patient developed decreased hand grip. Magnetic resonance imaging of the spine the next day showed a soft-tissue mass impinging on the spinal canal. The patient was subsequently taken to the operating room where the epidural abscess was drained.


Characterization of cadmium and calcium fluxes along the gut, malpighian tubules, and anal papillae of the dipteran Chironomus riparius.
Chironomids are often one of the dominant organisms in significantly polluted freshwater. Many invertebrate studies have characterized whole-organism mechanisms of toxicity, for example, assessing cadmium (Cd) uptake via calcium (Ca) channels. However, with the use of the scanning ion-selective electrode technique and an innovative Cd-selective microelectrode, we analyze this relationship at the organ level using a realistic concentration of Cd and Ca in the hemolymph (blood). Generally, Cd fluxes follow the same directional pattern as Ca, although Ca fluxes are approximately 5 times higher than those of Cd. These results correlate well with previous studies indicating that chironomids have a higher affinity for Ca over Cd, which affords them tolerance to Cd toxicity. When saline Ca concentration was increased to 10 times physiological levels, Cd fluxes from the gut lumen into the cells of the midgut regions were reduced by 50 to 80%. Transport of Cd from hemolymph to tissue for the posterior midgut, Malpighian tubule, and proximal ceca was also reduced by approximately 50%. The present results indicate that Cd fluxes into or across the gut and Malpighian tubules are reduced by high Ca, suggesting that Cd may be transported in some cells by similar mechanisms. However, Cd was actively excreted at the anal papillae after a 48-h waterborne exposure to Cd, but this process was independent of Ca and instead may involve a P-glycoprotein-related pump to detoxify Cd. Environ Toxicol Chem 2018;37:2542-2549. © 2018 SETAC.


Role of irradiation in management of synovial sarcoma: St. Jude Children's Research Hospital experience.
The role of irradiation in the management of synovial sarcoma (SS) in pediatric patients is evaluated. The review covers all children seen at St. Jude Children's Research Hospital between May 1969 and December 1992 with the diagnosis of soft tissue sarcoma, of the 37 patients with the subtype SS, 16 received irradiation for the management of primary site disease. There were four IRS Group I, six Group II, four Group III, and two Group IV patients receiving irradiation. Tumor grade included seven Grade II, and nine Grade III lesions. TMN staging identified eight T1 and eight T2 lesions. Follow-up has ranged from 14 to 117 months (med = 33 months). All IRS Group I patients had documented local control. Five of six IRS Group II and 4/4 Group III patients have had documented local control at last follow-up. IRS Group IV patients had either local control tumor stabilization (n=1) or evidence of tumor regression (n=1) at autopsy. Complications following irradiation include wound dehiscence (n=1), surgery to revise a painful scar (n=1) extremity length discrepancy (n=2), and femoral head avascular necrosis (n=1). At last follow-up, 10 of 14 patients receiving curative intent irradiation remain alive. This review indicates questionable benefit to the addition of irradiation for patients with adequate surgical resection and having "good" tumor characteristics (Grade I, II; IRS Group I, TMN T1A,T1B. For lesions that have had incomplete resection or partial response to chemotherapy, there is evidence that irradiation may provide durable local control. The role of irradiation in those patients with IRS Group IV disease is at present confined to palliative roles until the time when more effective chemotherapy will mandate the decision to treat primary disease for curative measures.


Adverse effects on Alfeios River Basin and an integrated management framework based on sustainability.
The Alfeios River, the longest and highest flow-rate river in Peloponnisos, constitutes an important water resource and ecosystem in Greece. In the present study, human activities in the Alfeios River Basin are described, and their impacts on water quality and the ecosystem are analyzed; effects resulting from interventions on river geomorphology between Flokas Dam and the river delta are determined. These actions have caused significant adverse impacts on the infrastructure (the dam, railroad, and road bridges), the level of aquifer water table and area water uses, and the aquatic and riparian ecosystem. A general integrated management strategy is formulated and a master management plan is proposed for resolving management problems in river basins. The plan considers local conditions and national requirements and complies with the European Communities legislation; it would help prevent further basin deterioration, improve water quality, and protect water resources and ecosystems in the area in accordance to sustainable development. The Alfeios River Basin serves as a case study in the development of the plan.


Amphiregulin mediates the hormonal regulation on Rspondin-1 expression in the mammary gland.
The steroid hormones are instrumental for the growth of mammary epithelial cells. Our previous study indicates that hormones regulate the expression of Rspondin-1 (Rspo1). Yet, the regulatory mechanism remains unknown. In the current study, we identify Amphiregulin (Areg) as a novel upstream regulator of Rspo1 expression mediating the hormonal influence. In response to hormonal signaling, Areg emanating from estrogen receptor (ER)-positive luminal cells, induce the expression of Rspo1 in ER-negative luminal cells. The paracrine action of Areg on Rspo1 expression is dependent on Egfr. Our data reveal a novel Estrogen-Areg-Rspo1 regulatory axis in the mammary gland, providing new evidence for the orchestrated action of systemic hormones and local growth factors.


Patient-perceived Burden of Disease in Pediatric Relapsing Polychondritis.
To assess patient-reported burden of disease in pediatric patients with relapsing polychondritis (RP) and to compare those findings to adult patients. A survey based on known clinical symptoms of RP was developed and administered to patients with a pediatric diagnosis of RP. Adult patients completed a similar survey. Twenty-one pediatric patients, or their parents, completed surveys. Median age at symptom onset was 6 years (interquartile range 1.8-12). Prior to diagnosis, most pediatric patients went to the emergency room (ER; 61.9%), saw > 3 physicians (57.1%), and took > 1 year to be diagnosed (61.9%). Pediatric patients were often diagnosed with asthma (42.9%), ear infections (42.9%), or sinusitis (33.3%) prior to diagnosis of RP. Symptoms prior to diagnosis included ear pain/redness (85.7%), joint pain/swelling (61.9%), and airway symptoms (38.1%). Four pediatric patients (19%) reported tracheomalacia requiring tracheostomy. Pediatric patients frequently missed school because of their disease (71.4%). Surveys from 290 adult patients were compared to pediatric patients. Pediatric patients were significantly more likely to undergo biopsy (42.9% vs 17.4%; p < 0.01) and be treated with biologics (42.9% vs 19%; p = 0.02). Adults were significantly more likely to be female (87.8% vs 28.6%; p < 0.01) and to report airway symptoms (77.9% vs 47.6%; p = 0.01). Prevalence of disease complications was not significantly different between adult and pediatric patients. The burden of disease in pediatric patients with RP includes missed school, diagnostic delay, ER visits, and multisystem disease, with resultant damage to cartilaginous structures. Differences in airway involvement and treatment approaches may exist between pediatric and adult patients.


Establishing a 'Virtual' model of emergency care in Melbourne's southeast.
Digital technologies, specifically those required to facilitate telehealth via an audiovisual medium, are now at a standard that allows them to reliably assess patients with acute complaints within their homes. The coronavirus disease 2019 (COVID-19) pandemic forced innovation and led to an increased acceptance of telehealth from both clinicians and patients and presented an opportunity to incorporate telehealth into emergency medicine practice. With inpatient capacity strain exacerbated by the pandemic, three hospital networks within Melbourne's geographical southeast collaborated with Ambulance Victoria (AV) to set up a virtual emergency department (VED) service in January 2022. This service aimed to allow certain patients to receive care in the more convenient setting of their homes. Referrals were made directly by AV personnel at the home while they were attending to these patients. Consultation with a VED clinician was initiated through a secure audiovisual platform, 'Healthdirect'. Following this consultation, care could be facilitated at home through the VED clinician providing advice regarding treatment that paramedics could administer or through linkage into one of the various outreach services provided by the organisations. The VED represents part of the initial integration of telehealth into our organisations. We hope our initial framework might be built upon through greater integration with community services and incorporating more digital technologies. The ongoing graded expansion of telehealth services within our organisations will likely see more and more patients managed in their own homes.


Microvessel quantification as a measure of angiogenic activity in benign breast-tissue lesions - a marker for precancerous disease.
Neovascularization is a key step in the transformation of normal breast tissue into a cancerous phenotype. We investigated the angiogenic activity of 58 cases of histologically confirmed benign breast lesions quantitating angiogenic activity by factor VIII highlighting of endothelial cells. The mean microvessel count (MVC) for all patients was 51 per 0.19 mm(2) in the area of highest angiogenic activity at x400 magnification (range 21-113 per mm(2)). MVC in benign tissues were similar to those found previously in cancer tissues. These data suggest that the 'angiogenic switch' occurs early in breast transformation and that a role may exist for the use of MVC in the identification of persons at heightened risk for subsequent development of breast cancer.


Young suicide attempters: a comparison between a clinical and an epidemiological sample.
To compare risk factors for self-harm in 2 groups: hospitalized adolescents who had attempted suicide and adolescents reporting suicide attempts in a community survey. All suicide attempters aged 13 to 19 years admitted to medical wards (n = 91) in a region of Norway were assessed and interviewed. Risk factors were identified by comparisons with a general population sample participating in a questionnaire study in the same community (n = 1,736). In this population sample, a separate analysis of risk factors for reporting deliberate self-harm (n = 141) was performed, applying bivariate and multivariate logistic regression models. Adjusted risk factors for suicide attempts in hospitalized adolescents were depression (odds ratio [OR] = 4.7), disruptive disorders (OR = 9.4), low self-worth (OR = 1.3), infrequent support from parents (OR = 3.3) or peers (OR = 3.3), parents' excessive drinking (OR = 4.3), and low socioeconomic status (OR = 2.4). For adolescents who self-reported self-harm, depression (OR = 3.1) and loneliness (OR = 1.13) were significant adjusted risk factors (p < .001). Low self-worth, low socioeconomic status, and little support from parents or peers characterized hospitalized suicidal adolescents compared with those who were not hospitalized. The risk factors were more powerful for hospitalized than for nonhospitalized adolescents. Prevention efforts should target the same factors for both groups, at a population level for nonhospitalized adolescents and at an individual level for hospitalized adolescents, with a focus on depression, low self-esteem, and family communication.


Needlestick safety and prevention act.
The Needlestick Safety and Prevention Act to revise the Occupational Safety and Health Administration's (OSHA) standard regulating occupational exposure to bloodborne pathogens, including the human immunodeficiency virus, the hepatitis B virus, and the hepatitis C virus, was signed into law on November 6, 2000. OSHA published in the Federal Register its regulations reflecting the Act and its requirments. The effective date of the regulations is April 18, 2001. The Needlestick Safety and Prevention Act seeks to further reduce health care workers' exposure to bloodborne pathogens by imposing additional requirements upon employers, such as hospitals and ASCs, concerning their sharps procedures. Consistent with the Act, OSHA's regulations (1) modify the definition of "engineering controls" and adds definitions for the terms "sharps with engineered sharps injury protection" and "needleless systems," (2) requires employers to consider and implement new technologies when they update their "exposure control plan," (3) requires employers to solicit employee input with respect to appropriate engineering controls, and (4) requires employers to maintain a sharps injury log. Practical questions about implementing the new requirements are a source of major concern. The House Committee on Education and the Workforce stated in legislative history to the Act that the statute was not meant to disturb the underlying flexible, performance-oriented nature of the Initial Standard.


Differential female sociality is linked with the fine-scale structure of sexual interactions in replicate groups of red junglefowl, Gallus gallus.
Recent work indicates that social structure has extensive implications for patterns of sexual selection and sexual conflict. However, little is known about the individual variation in social behaviours linking social structure to sexual interactions. Here, we use network analysis of replicate polygynandrous groups of red junglefowl (Gallus gallus) to show that the association between social structure and sexual interactions is underpinned by differential female sociality. Sexual dynamics are largely explained by a core group of highly social, younger females, which are more fecund and more polyandrous, and thus associated with more intense postcopulatory competition for males. By contrast, less fecund females from older cohorts, which tend to be socially dominant, avoid male sexual attention by clustering together and perching on branches, and preferentially reproduce with dominant males by more exclusively associating and mating with them. Collectively, these results indicate that individual females occupy subtly different social niches and demonstrate that female sociality can be an important factor underpinning the landscape of intrasexual competition and the emergent structure of animal societies.


Interaction of yeast repressor-activator protein Ume6p with glycogen synthase kinase 3 homolog Rim11p.
Meiosis and expression of early meiotic genes in the budding yeast Saccharomyces cerevisiae depend upon Rim11p, Ume6p, and Ime1p. Rim11p (also called Mds1p and ScGSK3) is a protein kinase related to glycogen synthase kinase 3 (GSK3); Ume6p is an architectural transcription factor; and Imelp is a Ume6p-binding protein that provides a transcriptional activation domain. Rim11p is required for Ime1p-Ume6p interaction, and prior studies have shown that Rim11p binds to and phosphorylates Ime1p. We show here that Rim11p binds to and phosphorylates Ume6p, as well. Amino acid substitutions in Ume6p that alter a consensus GSK3 site reduce or abolish Rim11p-Ume6p interaction and Rim11p-dependent phosphorylation, and they cause defects in interaction between Ume6p and Ime1p and in meiotic gene expression. Therefore, interaction between Rim11p and Ume6p, resulting in phosphorylation of Ume6p, is required for Ime1p-Ume6p complex formation. Rim11p, like metazoan GSK3beta, phosphorylates both interacting subunits of a target protein complex.


A comparative study of avermectin B1a and other modulators of the gamma-aminobutyric acid receptor . chloride ion channel complex.
The interactions of the anthelmintic agent avermectin B1a, the anticonvulsant pentobarbital, and the anxiolytic tracazolate with the gamma-aminobutyric acid (GABA) receptor . chloride ion channel complex in rat brain membrane were studied. The results indicated that they all potentiated ligand binding to the GABA and benzodiazepine receptors. The stimulatory effects of avermectin B1a and pentobarbital, but not tracazolate, on GABA receptor binding were inhibited by picrotoxin. The effect of avermectin B1a was not additive with those of tracazolate and pentobarbital. On the other hand, the stimulatory effect of GABA on benzodiazepine binding was additive with those of avermectin B1a and pentobarbital, but tracazolate and pentobarbital inhibited the effect of avermectin B1a. In the receptor heat inactivation experiments, avermectin B1a and clonazepam protected GABA receptors, whereas avermectin B1a and GABA protected benzodiazepine receptors. Tracazolate, pentobarbital, and picrotoxin did not protect either receptor. These findings suggest that the recognition sites for the benzodiazepines, avermectin B1a, pentobarbital, and picrotoxin are coupled allosterically to the GABA receptor . chloride ion channel complex in different ways. The binding sites for avermectin B1a may be partially shared by picrotoxin, pentobarbital, and tracazolate.


RanGTP-binding protein NXT1 facilitates nuclear export of different classes of RNA in vitro.
To better characterize the mechanisms responsible for RNA export from the nucleus, we developed an in vitro assay based on the use of permeabilized HeLa cells. This new assay supports nuclear export of U1 snRNA, tRNA, and mRNA in an energy- and Xenopus extract-dependent manner. U1 snRNA export requires a 5' monomethylated cap structure, the nuclear export signal receptor CRM1, and the small GTPase Ran. In contrast, mRNA export does not require the participation of CRM1. We show here that NXT1, an NTF2-related protein that binds directly to RanGTP, strongly stimulates export of U1 snRNA, tRNA, and mRNA. The ability of NXT1 to promote export is dependent on its capacity to bind RanGTP. These results support the emerging view that NXT1 is a general export factor, functioning on both CRM1-dependent and CRM1-independent pathways of RNA export.


Feline struvite urolithiasis: fasting reduced the effectiveness of a urinary acidifier (ammonium chloride) and increased the intake of a low magnesium diet.
In three separate experiments nine male cats were fed either a canned complete diet or a commercially available dry pelleted diet or the same dry diet containing 1.6 per cent ammonium chloride for seven days and then fasted for 20 hours. Then ad libitum feeding was continued and urine samples were taken at four-hour intervals for 12 hours and a final sample 12 hours later. Urine pH and the presence of struvite crystals in urine sediment were evaluated. The food and water intake of four of the nine cats was measured at the time of urine collection. After the fast, urine pH was raised, even after feeding the dry diet supplemented with ammonium chloride. A post prandial rise in urine pH was also seen on all three diets. After feeding the dry diet the postprandial peak pH was 7.97 and struvite occurred spontaneously. Urine pH after feeding the dry diet supplemented with ammonium chloride peaked at 7.75 then fell to 6.1 12 hours after the start of feeding. Struvite occurred spontaneously at all times until the pH reached 6.1 but when the pH of urine was raised to 7.0 the struvite crystallised. Urine pH on the canned complete diet peaked at 6.8 then fell to 5.8; struvite did not occur spontaneously but when urine pH was raised to 7.0 struvite crystallised except at the eighth and 12th hour sampling. These data show that fasting initiates a post prandial rise in urine pH and struvite crystalluria even when a normally effective urinary acidifier is used.(ABSTRACT TRUNCATED AT 250 WORDS)


Desulfobaculum xiamenensis gen. nov., sp. nov., a member of the family Desulfovibrionaceae isolated from marine mangrove sediment.
A taxonomic study was carried out on strain P1(T), which was isolated from mangrove sediment samples collected from Qinglan Port (Hainan, China). Cells were curved rods, that were motile, with a single polar flagellum. The strain was non-spore-forming with a cell size of 0.6×1.5-2.2 µm. Catalase and oxidase activities were not detected. Growth was observed in the temperature range 22-44 °C (optimum, 35-40 °C) and pH range 5.5-8.5 (optimum, pH 7.0). NaCl was required for growth and tolerated at up to 3.5% (w/v) (optimum, 0.5%). Strain P1(T) utilized hydrogen, succinate, L-malate, citrate, oxalate, DL-lactate, pyruvate, or cysteine as electron donors, and sulfate or sulfite as electron acceptors. Fermentation products from pyruvate were acetate, H(2) and CO(2). Phylogenetic analyses based on 16S rRNA gene sequences showed that strain P1(T) formed a distinct evolutionary lineage within the family Desulfovibrionaceae. Strain P1(T) was most closely related to members of the genera Desulfovibrio (92.0-94.3% 16S rRNA gene sequence similarity), Desulfocurvus (91.1%), Bilophila (87.9%) and Lawsonia (86.0%) of the family Desulfovibrionaceae. The DNA G+C content of strain P1(T) was 64.5 mol% and the major cellular fatty acids were iso-C(15:0) (18.8%), anteiso-C(15:0) (5.0%), C(16:0) (14.2%) and iso-C(17:1)ω9c (24.4%). The predominant menaquinone was MK-7 (97%). Major polar lipids were phosphatidylcholine, phosphatidylethanolamine and phosphatidylglycerol. Strain P1(T) was distinguishable from members of phylogenetically related genera by differences in several phenotypic properties. On the basis of the phenotypic and phylogenetic data, strain P1(T) represents a novel species of a new genus, for which the name Desulfobaculum xiamenensis gen. nov., sp. nov. is proposed. The type strain of Desulfobaculum xiamenensis is P1(T) (=CGMCC 1.5166(T)=DSM 24233(T)).


Hypoxic coordinate regulation of mitochondrial enzymes in mammalian cells.
The effect of hypoxic exposure on various mitochondrial enzymes and on cell mitochondrial genomic content was studied in two types of mammalian cells. Hypoxia depressed the activity of six enzymes to the same degree. The kinetics of depression and of recovery during reexposure to normoxia were statistically similar for three marker enzymes. Despite the global and symmetrical decrease in enzyme activities, mitochondrial DNA remained constant. This suggests either symmetrical loss of mitochondrial enzymes from all mitochondria or complete loss of enzymes from a subpopulation of mitochondria with retention of an intact mitochondrial genome.


A simple assay for inhibitors of mycobacterial oxidative phosphorylation.
Oxidative phosphorylation, the combined activities of the electron transport chain (ETC) and adenosine triphosphate (ATP) synthase, has emerged as a valuable target for antibiotics to treat infection with Mycobacterium tuberculosis and related pathogens. In oxidative phosphorylation, the ETC establishes a transmembrane electrochemical proton gradient that powers ATP synthesis. Monitoring oxidative phosphorylation with luciferase-based detection of ATP synthesis or measurement of oxygen consumption can be technically challenging and expensive. These limitations reduce the utility of these methods for characterization of mycobacterial oxidative phosphorylation inhibitors. Here we show that fluorescence-based measurement of acidification of inverted membrane vesicles (IMVs) can detect and distinguish between inhibition of the ETC, inhibition of ATP synthase, and non-specific membrane uncoupling. In this assay, IMVs from M. smegmatis are acidified either through the activity of the ETC or ATP synthase, the latter modified genetically to allow it to serve as an ATP-driven proton pump. Acidification is monitored by fluorescence from 9-amino-6-chloro-2-methoxyacridine, which accumulates and quenches in acidified IMVs. Non-specific membrane uncouplers prevent both succinate- and ATP-driven IMV acidification. In contrast, the ETC Complex III2IV2 inhibitor telacebec (Q203) prevents succinate-driven acidification but not ATP-driven acidification and the ATP synthase inhibitor bedaquiline prevents ATP-driven acidification but not succinate-driven acidification. We use the assay to show that, as proposed previously, lansoprazole sulfide is an inhibitor of Complex III2IV2 while thioridazine uncouples the mycobacterial membrane non-specifically. Overall, the assay is simple, low cost, and scalable, which will make it useful for identifying and characterizing new mycobacterial oxidative phosphorylation inhibitors.


Hydroxyapatite eye implant.
Hydroxyapatite, a coral, provides an effective foundation for a prosthetic eye. With the implant, the prosthetic eye has a normal appearance, including movement. The infusion of blood into the implant prevents it from behaving as a foreign object in the patient's body.


Flow cytometry for the analysis of α-dystroglycan glycosylation in fibroblasts from patients with dystroglycanopathies.
α-dystroglycan (α-DG) is a peripheral membrane protein that is an integral component of the dystrophin-glycoprotein complex. In an inherited subset of muscular dystrophies known as dystroglycanopathies, α-DG has reduced glycosylation which results in lower affinity binding to several extracellular matrix proteins including laminins. The glycosylation status of α-DG is normally assessed by the binding of the α-DG antibody IIH6 to a specific glycan epitope on α-DG involved in laminin binding. Immunocytochemistry and immunoblotting are two of the most widely used methods to detect the amount of α-DG glycosylation in muscle. While the interpretation of the presence or absence of the epitope on muscle using these techniques is straightforward, the assessment of a mild defect can be challenging. In this study, flow cytometry was used to compare the amount of IIH6-reactive glycans in fibroblasts from dystroglycanopathy patients with defects in genes known to cause α-DG hypoglycosylation to the amount in fibroblasts from healthy and pathological control subjects. A total of twenty one dystroglycanopathy patient fibroblasts were assessed, as well as fibroblasts from three healthy controls and seven pathological controls. Control fibroblasts have clearly detectable amounts of IIH6-reactive glycans, and there is a significant difference in the amount of this glycosylation, as measured by the mean fluorescence intensity of an antibody recognising the epitope and the percentage of cells positive for the epitope, between these controls and dystroglycanopathy patient fibroblasts (p<0.0001 for both). Our results indicate that the amount of α-DG glycosylation in patient fibroblasts is comparable to that in patient skeletal muscle. This method could complement existing immunohistochemical assays in skeletal muscle as it is quantitative and simple to perform, and could be used when a muscle biopsy is not available. This test could also be used to assess the pathogenicity of variants of unknown significance in genes involved in dystroglycanopathies.


Muscle cell identity requires Pax7-mediated lineage-specific DNA demethylation.
Skeletal muscle stem cells enable the formation, growth, maintenance, and regeneration of skeletal muscle throughout life. The regeneration process is compromised in several pathological conditions, and muscle progenitors derived from pluripotent stem cells have been suggested as a potential therapeutic source for tissue replacement. DNA methylation is an important epigenetic mechanism in the setting and maintenance of cellular identity, but its role in stem cell determination towards the myogenic lineage is unknown. Here we addressed the DNA methylation dynamics of the major genes orchestrating the myogenic determination and differentiation programs in embryonic stem (ES) cells, their Pax7-induced myogenic derivatives, and muscle stem cells in proliferating and differentiating conditions. Our data showed a common muscle-specific DNA demethylation signature required to acquire and maintain the muscle-cell identity. This specific-DNA demethylation is Pax7-mediated, and it is a prime event in muscle stem cells gene activation. Notably, downregulation of the demethylation-related enzyme Apobec2 in ES-derived myogenic precursors reduced myogenin-associated DNA demethylation and dramatically impaired the expression of differentiation markers and, ultimately, muscle differentiation. Our results underscore DNA demethylation as a key mechanism driving myogenesis and identify specific Pax7-mediated DNA demethylation signatures to acquire and maintain the muscle-cell identity. Additionally, we provide a panel of epigenetic markers for the efficient and safe generation of ES- and induced pluripotent stem cell (iPS)-derived myogenic progenitors for therapeutic applications.


Risk of homelessness after prison release and recidivism in Denmark: a nationwide, register-based cohort study.
Transitional periods between and across services have been linked to homelessness. We aimed to investigate the association of previous history of homelessness and psychiatric disorders with risk of homelessness after release from prison. Additionally, we examined the association between homelessness after release and risk of recidivism. We did a nationwide, register-based cohort study of people aged 15 years or older who were released from prison for the first time in Denmark between Jan 1, 2001, and Dec 31, 2021. We obtained data using the Danish Civil Registration System with data linked across other registries (the Danish Central Criminal Register, the Danish Homeless Register, the Danish National Patient Register, and the Danish Psychiatric Central Research Register) on release date, homeless shelter contacts, psychiatric disorders, and new convictions. Outcomes were homelessness after release from prison, defined as first homeless shelter contact following release from first imprisonment, and recidivism within 2 years of release, defined as the first police-recorded criminal conviction after prison release. We calculated incidence rates per 1000 person-years, incidence rate ratios (IRRs) using Poisson regression analysis, and probability of homelessness and recidivism after release. Sex, age, calendar year, country of origin, highest educational level, relationship status, and length of index imprisonment were included as confounders. The study cohort included 37 382 individuals (34 792 males [93·1%] and 2590 females [6·9%]) aged 15-41 years, who were released from prison between Jan 1, 2001, and Dec 31, 2021, contributing 202 197 person-years at risk. Mean follow-up duration was 5·4 person-years (SD 5·6). Overall, 1843 (4·9%) of 37 382 individuals became homeless. 1 year after release from prison, 788 (2·1%) of 37 382 individuals had at least one homeless shelter contact, and among 1761 individuals with previous history of homelessness before index imprisonment, 357 (20·7%) became homeless. The incidence of homelessness after release was 102·5 cases per 1000 person-years for individuals with previous history of homelessness and 6·7 cases per 1000 person-years in individuals without (IRR 16·4, 95% CI 14·8-18·2; adjusted for sex, age, and calendar year). Individuals who additionally had a mental illness had a higher risk of homelessness (IRR 22·6, 19·7-25·9) compared with those without either previous homelessness or mental illness, and a substantially higher risk was observed for those with previous homelessness and drug use disorder (25·0, 21·6-28·9) compared with those without. Within 2 years of release from prison, the probability of recidivism was 73·2% (95% CI 72·8-73·7). The risk of recidivism was higher among people experiencing homelessness after release from prison than those who did not experience homelessness after release (IRR 1·5, 95% CI 1·3-1·7), adjusted for sex, age, and calendar year. Criminal justice services should review approaches to reduce risk of homelessness, and consider improving liaison with mental health and substance misuse services to prevent adverse outcomes on release from prison. Clinical guidelines applied to criminal justice settings should address the health of individuals who experience homelessness. Lundbeck Foundation.


Purification and Characterization of an Extracellular T2 Family Ribonuclease (RNase) from Bacillus megaterium NRRL 3712.
Ribonucleases of T2 family are ubiquitous cellular components which have played several biological functions in molecular and pharmaceutical fields. Therefore, a soluble and highly active RNase belonging to T2 family was screened from Bacillus megaterium NRRL 3712, and different cultivation strategies were applied to enhance the production of enzyme. A high-level of an extracellular RNase and cell density was produced using optimal cultivation conditions. A monomeric enzyme with a molecular mass of 45 kDa, was purified to homogeneity using acetone precipitation and ion-exchange chromatography. Purified enzyme was optimally activity at 45°C and pH 7.0, and it displayed a half-life of 26 min at 64°C. It was quite stable up to 60 min at 40-50°C temperature and over a broad range of pH 4.5-8.0. It showed great substrate specificity with yeast RNA, poly (A), poly (G), poly (C), and poly (U). Kinetic parameters such as Km, Vmax, kcat and kcat Km -1 values against yeast RNA as substrate, were 71.67 µg mL-1, 7866.4 μmol mg-1min-1, 17669.4 sec-1, and 246.53, respectively. The article provides a valuable novel RNase which exhibited great resistance against various organic solvents, detergents and metal ions, whereas its activity was stimulated up to 142% by adding 5 mM EDTA. Hence, dictates its applicability as therapeutic agent and in various other biotechnological fields.


External detection of early microvascular dysfunction after no-flow ischemia followed by reperfusion in isolated rabbit hearts.
To define relationships better between the duration of severe ischemia and microvascular functional integrity with an approach potentially applicable to studies in vivo, the effects of 30 and 60 minutes of global, no-flow ischemia on the coronary vasculature of isolated, perfused rabbit hearts were determined. Residue-detection data, analyzed with a two-compartment model, were used to estimate indices of microvascular function, including the mean-transit time (tBSA) of radiolabeled bovine serum albumin (125I-BSA), vascular into extravascular space clearance, and vascular and extravascular space volumes. It was shown that the Central Volume Principle of tracer kinetics does not hold when transport of label between vascular and extravascular spaces takes place convectively by solvent drag, and a more general expression for tBSA was derived and applied. Left ventricular end-diastolic pressure and left ventricular developed pressure were monitored with an isovolumic balloon. Aortic perfusion pressure, left ventricular end-diastolic pressure, left ventricular developed pressure and vascular space volume remained constant, while mean transit time, vascular into extravascular space clearance and extravascular space volumes increased gradually during 3-hour control perfusions. Perfusion pressure, mean transit time and extravascular space clearance increased significantly with reperfusion after 30 minutes of ischemia even though left ventricular end-diastolic and left ventricular-developed pressures returned to control levels. Vascular space volumes increased minimally, whereas extravascular space volumes increased 5-fold during reperfusion. These changes in 125I-BSA washout and permeation across endothelium with reperfusion after no-flow ischemia indicate that compromised vascular integrity is an early manifestation of ischemia with functional consequences that persist even after ischemia sufficiently brief to permit restoration of left ventricular performance.


Ribociclib: First Global Approval.
Ribociclib is an oral, small-molecule inhibitor of cyclin-dependent kinase (CDK) 4 and 6 that is under development by Novartis for the treatment of cancer. CDKs play an important role in cell cycle progression and cellular proliferation, and inhibition of these kinases with ribociclib results in G1 phase cell-cycle arrest. Ribociclib, in combination with an aromatase inhibitor, was recently approved in the USA for the first-line treatment of advanced breast cancer and has been submitted for approval in the EU for this indication. Ribociclib is undergoing further phase III investigations in breast cancer and is being evaluated in phase I or II trials for various solid tumour types and haematological malignancies. This article summarizes the milestones in the development of ribociclib leading to this first global approval for use as initial endocrine-based therapy for the treatment of postmenopausal women with hormone receptor-positive, human epidermal growth factor receptor 2-negative advanced or metastatic breast cancer.


Insights Gained from RNA Editing Targeted by the CRISPR-Cas13 Family.
Clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein (Cas) systems, especially type II (Cas9) systems, have been widely developed for DNA targeting and formed a set of mature precision gene-editing systems. However, the basic research and application of the CRISPR-Cas system in RNA is still in its early stages. Recently, the discovery of the CRISPR-Cas13 type VI system has provided the possibility for the expansion of RNA targeting technology, which has broad application prospects. Most type VI Cas13 effectors have dinuclease activity that catalyzes pre-crRNA into mature crRNA and produces strong RNA cleavage activity. Cas13 can specifically recognize targeted RNA fragments to activate the Cas13/crRNA complex for collateral cleavage activity. To date, the Cas13X protein is the smallest effector of the Cas13 family, with 775 amino acids, which is a promising platform for RNA targeting due to its lack of protospacer flanking sequence (PFS) restrictions, ease of packaging, and absence of permanent damage. This study highlighted the latest progress in RNA editing targeted by the CRISPR-Cas13 family, and discussed the application of Cas13 in basic research, nucleic acid diagnosis, nucleic acid tracking, and genetic disease treatment. Furthermore, we clarified the structure of the Cas13 protein family and their molecular mechanism, and proposed a future vision of RNA editing targeted by the CRISPR-Cas13 family.


Detection of flavokavins (A, B, C) in cultivars of kava (Piper methysticum) using high performance thin layer chromatography (HPTLC).
Kava (Piper methysticum) is used to prepare the traditional beverage of the Pacific islands. In Europe, kava has been suspected to cause hepatoxicity with flavokavin B (FKB) considered as a possible factor. The present study describes an HPTLC protocol for rapid screening of samples. The objectives are: to detect the presence of flavokavins in extracts and to compare the FKB levels in different cultivars. Overall, 172 samples originating from four cultivars groups (noble, medicinal, two-days and wichmannii), were analysed. Results indicate that the ratio FKB/kavalactones is much higher in two-days (0.39) and wichmannii (0.32) compared to nobles (0.09) and medicinal cultivars (0.10). For each group, the ratios flavokavins/kavalactones do not change significantly between roots, stumps or basal stems and among clones, indicating that they are genetically controlled. This protocol has good accuracy and is cost efficient for routine analysis. We discuss how it could be used for quality control.


Disruption of a rice pentatricopeptide repeat protein causes a seedling-specific albino phenotype and its utilization to enhance seed purity in hybrid rice production.
The pentatricopeptide repeat (PPR) gene family represents one of the largest gene families in higher plants. Accumulating data suggest that PPR proteins play a central and broad role in modulating the expression of organellar genes in plants. Here we report a rice (Oryza sativa) mutant named young seedling albino (ysa) derived from the rice thermo/photoperiod-sensitive genic male-sterile line Pei'ai64S, which is a leading male-sterile line for commercial two-line hybrid rice production. The ysa mutant develops albino leaves before the three-leaf stage, but the mutant gradually turns green and recovers to normal green at the six-leaf stage. Further investigation showed that the change in leaf color in ysa mutant is associated with changes in chlorophyll content and chloroplast development. Map-based cloning revealed that YSA encodes a PPR protein with 16 tandem PPR motifs. YSA is highly expressed in young leaves and stems, and its expression level is regulated by light. We showed that the ysa mutation has no apparent negative effects on several important agronomic traits, such as fertility, stigma extrusion rate, selfed seed-setting rate, hybrid seed-setting rate, and yield heterosis under normal growth conditions. We further demonstrated that ysa can be used as an early marker for efficient identification and elimination of false hybrids in commercial hybrid rice production, resulting in yield increases by up to approximately 537 kg ha(-1).


Homogenization-resistant and -susceptible components of tobacco mosaic virus replicative form RNA.
When prepared from tissue frozen with liquid nitrogen, tobacco mosaic virus replicative form RNA (TMV RF) was uniform in size but when prepared by high-speed homogenization, or when TMV RF prepared with liquid nitrogen was homogenized, 80 to 90% of the RF broke into relatively discrete pieces. The unbroken RF was not fragmented by additional homogenization. The TMV RF components susceptible and resistant to breakage, respectively, were synthesized with similar kinetics in relation to length of labelling period, but the slightly more resistant component was synthesized during the early infection period. Both components were produced by different strains of TMV but leaves infected with cowpea chlorotic mottle or southern bean mosaic viruses yielded only RF resistant to breakage. TMV replicative intermediate RNA was also broken by homogenization. The occurrence of the two RF components may be of significance in the replication of RNA viruses.


Relationship between protein phosphatase type-2C activity and induction of apoptosis in cultured neuronal cells.
The cellular composition and concentration of fatty acids are crucial for proliferation and survival. We recently showed stimulation of protein phosphatase type-2C (PP2C) by unsaturated fatty acids. Here, we describe that treatment of cultured chick neurons with 100 microM oleic acid for 24h increased the percentage of damaged neurons to 61+/-9% compared with 25+/-4% in controls. Oleic acid-induced cell death showed features of apoptosis such as chromatin condensation, shrinkage of the nucleus, DNA fragmentation and caspase-3 activation. Extensive studies with a variety of fatty acids revealed a striking correlation between activation of PP2C and induction of apoptosis. Lipophilicity, oxidizability, and an acidic group were required for both effects. In addition, activation of PP2C and induction of apoptosis could discriminate between cis- and trans-conformation of the fatty acids. The results are in favor of PP2C playing an important, yet unidentified role in apoptosis.


Comparison of conventional and organic management conditions on growth performance, carcass characteristics and haematological parameters in Karacabey Merino and Kivircik breeds.
Growth performance, carcass characteristics, post-slaughtering and haematological parameters of Kivircik and Karacabey Merino male lambs in conventional and organic management systems were compared. The animals which were weaned at 7 weeks of age were divided into Kivircik conventional, Kivircik organic (KO), Karacabey Merino conventional and Karacabey Merino organic (MO) groups containing 12 lambs each. Fattening was ended when lambs attained 35 kg of live weight. The time to attain the determined fattening weight was significantly different among the groups, and Merino lambs having higher live weight gain were earlier than Kivircik lambs (p < 0.05). Overall conventional (CG) and organic group lambs were also compared. Live weight gain, intra-abdominal fat amount, external fat thickness and visceral organ weight were significantly higher in CG lambs (p < 0.05). Higher haematocrit and erythrocyte counts were obtained with the CG group (p < 0.05), whilst triglyceride, total plasma cholesterol and lipoprotein (HDL, LDL, VLDL) levels between groups were not significant. Pneumonia was the unique infection, with an incidence of 50% (six lambs) and 16.6% (two lambs) for MO and KO animals, respectively. The mortality rate was 16.6% (two lambs) for MO group, whilst no mortality was recorded for KO group animals. The present study has shown that although Karacabey merino lambs had higher growth performance compared to Kivircik lambs, organically fattened lambs in whole exhibited inferior growth performance. Lower infection and mortality observed with Kivircik lambs suggested that they could be more resistant to infections and outdoor environmental conditions.


Cell-autonomous defense, re-organization and trafficking of membranes in plant-microbe interactions.
Plant cells dynamically change their architecture and molecular composition following encounters with beneficial or parasitic microbes, a process referred to as host cell reprogramming. Cell-autonomous defense reactions are typically polarized to the plant cell periphery underneath microbial contact sites, including de novo cell wall biosynthesis. Alternatively, host cell reprogramming converges in the biogenesis of membrane-enveloped compartments for accommodation of beneficial bacteria or invasive infection structures of filamentous microbes. Recent advances have revealed that, in response to microbial encounters, plasma membrane symmetry is broken, membrane tethering and SNARE complexes are recruited, lipid composition changes and plasma membrane-to-cytoskeleton signaling is activated, either for pre-invasive defense or for microbial entry. We provide a critical appraisal on recent studies with a focus on how plant cells re-structure membranes and the associated cytoskeleton in interactions with microbial pathogens, nitrogen-fixing rhizobia and mycorrhiza fungi.


[Circumcision--criticism of the routine].
Circumcision is one of the most frequent operative procedures done in males. About 120 circumcisions are performed every 5 minutes over the world [14]. Three different reasons lead to circumcision: 1) Medical reasons in present of a pathologic phimosis. 2) Circumcisions done due to religious, social or cultural rea-reasons. 3) Finally in many countries circumcision is performed as "routine-circumcision" in the newborn period. While in the United States the number of routine-circumcisions decreases (about 60% of all male newborns) South-Korea has a rate near to 100%. Even with no religious or cultural background in Germany circumcision often is performed without scrutinizing medical indication. Circumcision is regarded as an procedure with no complications and no disadvantage for the patient. In general circumcision has no medical benefit neither in decreasing the incidence of urinary tract infections nor of sexual transmitted diseases nor of neoplasias. Medical indication for circumcision is given in present of pathologic phimosis in 4% of all males. Postoperative complications range up to 2% and "circumcision is the amputation of the prepuce from the rest of the penis, resulting in permanent alteration of the anatomy, histology and function of the penis...". There are many reports about having discomfort and disadvantages after circumcision as well to the males as to the sexual partners. This challenge the legality of neonatal involuntary circumcision because legality is based on saving the children's best interests.


[Perception of HIV infection prevention messages by truck drivers in Togo with respect to the <<Roulez Protégé>> billboard].
Togo is one of the West African countries in which HIV prevalence remains high in the general population. Several HIV prevention interventions have targeted truck drivers. The purpose of this study was to describe and analyse the perceptions of truck drivers with respect to the HIVprevention message of the <<Roulez Protégé>> billboard. A qualitative study was conducted among truck drivers at "Terminal du Sahel" in Lomé in May 2013 in French and sometimes in a local language. The data generated by this survey were submitted to qualitative analysis. A total of 24 truck drivers were interviewed. They had already heard about HIV/AIDS and were able to list various modes of HIV transmission and various ways of protecting oneselffrom HIV However, they had a poor perception of the risk of contracting HIV infection. Although all participants had seen the "Roulez Protégé" billboard several times in various places, it made them feelguilty, as itsuggested that they were responsiblefor spread of the HIV/AIDS epidemic. Truck drivers had a poor understanding of the message expressed by this billboard. In Togo, truck drivers constitute a group at high risk of HIV in which prevention interventions must be intensified.


Stimulus-response coupling in monocytes infected with Leishmania. Attenuation of calcium transients is related to defective agonist-induced accumulation of inositol phosphates.
Mononuclear phagocytes infected with Leishmania have been shown to have defective responses to extracellular stimuli. To investigate the potential relationship of these findings to alterations in calcium-dependent signaling pathways, the regulation of [Ca2+]i concentrations was examined in human peripheral blood monocytes infected with amastigotes of Leishmania donovani. Measurements of [Ca2+]i in fura-2-loaded monocytes were made at the single cell level by microfluorimetry. In normal monocytes, resting [Ca2+]i was 56 +/- 2 nM (mean +/- SEM). In contrast, in monocytes infected with Leishmania there was an approximately twofold increase in basal [Ca2+]i (122 +/- 5 nM, p less than 0.01 vs control). Treatment of cells with pertussis toxin before infection did not abrogate infection-induced increases in basal [Ca2+]i, suggesting that this effect was not mediated via the activation of a G protein coupled to phospholipase C. However, elevated resting [Ca2+]i did correlate with increased rates of 45Ca2+ uptake by infected monocytes. As expected, in response to treatment with 10(-7) M FMLP, control monocytes showed rapid net increases in [Ca2+]i of 303 +/- 19 nM. In contrast, net transients of [Ca2+]i in infected monocytes in response to FMLP were attenuated to only 137 +/- 9 nM (p less than 0.01 vs control). This result was not related to excess buffering of [Ca2+]i in infected cells as both control and infected monocytes showed equivalent transients of [Ca2+]i in response to the calcium ionophore A23187. Rather, inhibition of agonist-induced calcium release in infected cells appeared related to defective generation of second messenger because compared to control cells labeled with myo-[2-3H]inositol, little accumulation of inositol 1,4,5-trisphosphate was detected in infected monocytes. Attenuation of inositol phosphate accumulation and calcium release in response to chemotactic peptide correlated with decreased FMLP-induced superoxide and hydrogen peroxide production by infected monocytes. These results provide direct evidence for defective regulation of [Ca2+]i and calcium-dependent signaling in Leishmania-infected monocytes and provide a basis for understanding abnormalities in activation-related responses that involve signaling through Ca(2+)-regulated pathways.


Short-term UV exposure of sunbathers at a Mediterranean Sea site.
The aim of this study was to investigate whether systematic differences in solar UV exposure on a specific anatomical site (chest) exist among three groups of Italian sunbathers: healthy subjects (suntanned and non-suntanned individuals) and subjects affected by abnormally high sensitivity to solar exposure. A second aim of the study was to search for a possible relation among biological markers of individual response to UV exposure (such as skin colorimetric parameters, skin temperature and changes in free radical amounts [FR] in the blood) and photosensitivity. FR in the blood were analyzed because of their possible influence on UV carcinogenesis. Measurements of ambient doses (i.e. incident erythemally weighted irradiance on a horizontal surface over a specified period of time) and erythemally effective UV dose received by an anatomical site (here called personal dose or exposure on a specific anatomical site) were investigated. Personal doses received by the chest were determined using polysulfone dosimetry. Exposure Ratio (ER), defined as the ratio between the personal dose and the corresponding ambient dose during the same exposure period, was then calculated. Measuring of skin color in the Commission Internationale de l'Eclairage L (luminance), a (redness), b (yellowness) system and skin temperature were also carried out on the inner upper arm (nonexposed skin site) and on the chest. It was found that the median value of ER was 0.20 (min: 0.09 and max: 0.34) for suntanned individuals, it was 0.17 (min: 0.13 and max: 0.42) for non-suntanned individuals and it was 0.19 (min: 0.14 and max: 0.34) for photosensitive individuals. There were no significant differences across the groups in their median ER scores. In addition, the statistical analysis showed that L on the exposed site before exposure demonstrated consistently higher median scores after exposure in all groups. The b value after exposure was significantly lower than before exposure in all participants, while no significant differences for a were observed before or after exposure between or within the groups. Our findings suggest that photodermatoses are not significantly related to ER and to the changes in biological markers due to too short-term UV exposure.


Activity Subspaces in Medial Prefrontal Cortex Distinguish States of the World.
Medial prefrontal cortex (mPfC) activity represents information about the state of the world, including present behavior, such as decisions, and the immediate past, such as short-term memory. Unknown is whether information about different states of the world are represented in the same mPfC neural population and, if so, how they are kept distinct. To address this, we analyze here mPfC population activity of male rats learning rules in a Y-maze, with self-initiated choice trials to an arm end followed by a self-paced return during the intertrial interval (ITI). We find that trial and ITI population activity from the same population fall into different low-dimensional subspaces. These subspaces encode different states of the world: multiple features of the task can be decoded from both trial and ITI activity, but the decoding axes for the same feature are roughly orthogonal between the two task phases, and the decodings are predominantly of features of the present during the trial but features of the preceding trial during the ITI. These subspace distinctions are carried forward into sleep, where population activity is preferentially reactivated in post-training sleep but differently for activity from the trial and ITI subspaces. Our results suggest that the problem of interference when representing different states of the world is solved in mPfC by population activity occupying different subspaces for the world states, which can be independently decoded by downstream targets and independently addressed by upstream inputs.SIGNIFICANCE STATEMENT Activity in the medial prefrontal cortex plays a role in representing the current and past states of the world. We show that during a maze task, the activity of a single population in medial prefrontal cortex represents at least two different states of the world. These representations were sequential and sufficiently distinct that a downstream population could separately read out either state from that activity. Moreover, the activity representing different states is differently reactivated in sleep. Different world states can thus be represented in the same medial prefrontal cortex population but in such a way that prevents potentially catastrophic interference between them.


Factor score estimation in multimethod measurement designs with planned missing data.
Multimethod measurement designs with planned missing data (MMM-PMD) aim at combining cheap proxy methods (e.g., self-reports) with an expensive gold standard method (e.g., biomarker) in order to improve the cost-efficiency of research designs. This article presents a comprehensive simulation study investigating whether accurate factor scores with trustworthy confidence or credible intervals for the gold standard method can be obtained in MMM-PMD designs. The results indicate that the Bartlett-FIML, regression-FIML, and fully Bayesian estimator perform equally well in terms of recovering the true rank-order and absolute level of the factor scores for subjects with complete data. However, for subjects with planned missing data the estimated factor scores are considerably biased and cannot be computed with the Bartlett-FIML estimator. The confidence or credible intervals for the estimated factor scores are trustworthy for complete cases. For subjects with planned missing data, the coverage rates vary considerably across conditions. Thus, the present study illustrates that individual scores from MMM-PMD designs should not be used for consequential individual decisions. Still, factor scores from MMM-PMD designs might be useful when used for secondary analyses that rely on group-level statistics like computing propensity scores for matching purposes or for factor score regression. The simulation results underline that multiple reliable proxy methods that are highly correlated with the gold standard method but are not highly correlated with each other should be administered to maximize the accuracy of the estimated factor scores as well as the trustworthiness of the corresponding confidence or credible intervals. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


2-Benzoyl-4-chloro-phenyl benzoate.
In the title compound, C20H13ClO3, the dihedral angles between the benzoate and the chloro-benzene and benzoyl rings are 68.82 (5) and 53.76 (6)°, respectively, while the dihedral angle between the benzoyl and benzoate rings is 81.17 (5)°. The eight atoms of the benzoyl residue are essentially planar with the exception of the O atom which lies 0.1860 (5) Å out of their mean plane (r.m.s. deviation = 0.97 Å). The nine atoms of benzoate residue are also essentially planar (r.m.s. deviation = 0.20 Å) with the ester O atom showing the greatest deviation [0.407 (12) Å] from their mean plane. In the crystal, mol-ecules are connected into centrosymmetric dimers by pairs of C-H⋯O hydrogen bonds.


bcr/abl mRNA detection following bone marrow transplantation for chronic myelogenous leukemia.
We have utilized the polymerase chain reaction (PCR) to sensitively detect persistence of the chronic myelogenous leukemia (CML) malignant clone and to study bcr/abl mRNA splicing patterns following bone marrow transplantation. Thirteen of sixteen patients displayed persistent malignant cells during post-BMT clinical remission. In two patients bcr/abl mRNA was detected 4 and 9 months prior to clinical relapse. In eleven of fourteen patients in continued clinical remission malignant cells were detected post-BMT. Ten of these eleven patients were also cytogenetically normal. Seven patients have lost all evidence of bcr/abl transcript, but only at 1-2 years posttransplant, while four have shown persistence of the bcr/abl transcript from 28 days to 3 years post-BMT and one has converted from an initially negative result at 1 year post-BMT to detectable levels of chimeric mRNA at 2 years. Thus, 8/9 patients tested at or before 6 months, 7/12 at 1 year, and 3/10 at 2 years showed persistent detectable CML cells. Intriguingly, mRNA splicing patterns changed in 5 patients following BMT, with complete loss of mRNA containing bcr exon 3 (n = 2) or new appearance of mRNA not containing bcr exon 3 (n = 2). A single patient transiently lost evidence of bcr exon 3 expression while persistently expressing the bcr exon 2/abl exon 2 splice. Our data suggest that the majority of patients harbor small numbers of malignant cells following transplantation, and that such persistence may not inevitably predict clinical relapse. Complete elimination of the malignant CML clone post-BMT may rely on immunological mechanisms (e.g., graft-vs-leukemia).


Interleukin 1 receptor antagonist knockout mice show enhanced microglial activation and neuronal damage induced by intracerebroventricular infusion of human beta-amyloid.
Interleukin 1 (IL-1) is a key mediator of immune responses in health and disease. Although classically the function of IL-1 has been studied in the systemic immune system, research in the past decade has revealed analogous roles in the CNS where the cytokine can contribute to the neuroinflammation and neuropathology seen in a number of neurodegenerative diseases. In Alzheimer's disease (AD), for example, pre-clinical and clinical studies have implicated IL-1 in the progression of a pathologic, glia-mediated pro-inflammatory state in the CNS. The glia-driven neuroinflammation can lead to neuronal damage, which, in turn, stimulates further glia activation, potentially propagating a detrimental cycle that contributes to progression of pathology. A prediction of this neuroinflammation hypothesis is that increased IL-1 signaling in vivo would correlate with increased severity of AD-relevant neuroinflammation and neuronal damage. To test the hypothesis that increased IL-1 signaling predisposes animals to beta-amyloid (Abeta)-induced damage, we used IL-1 receptor antagonist Knock-Out (IL1raKO) and wild-type (WT) littermate mice in a model that involves intracerebroventricular infusion of human oligomeric Abeta1-42. This model mimics many features of AD, including robust neuroinflammation, Abeta plaques, synaptic damage and neuronal loss in the hippocampus. IL1raKO and WT mice were infused with Abeta for 28 days, sacrificed at 42 days, and hippocampal endpoints analyzed. IL1raKO mice showed increased vulnerability to Abeta-induced neuropathology relative to their WT counterparts. Specifically, IL1raKO mice exhibited increased mortality, enhanced microglial activation and neuroinflammation, and more pronounced loss of synaptic markers. Interestingly, Abeta-induced astrocyte responses were not significantly different between WT and IL1raKO mice, suggesting that enhanced IL-1 signaling predominately affects microglia. Our data are consistent with the neuroinflammation hypothesis whereby increased IL-1 signaling in AD enhances glia activation and leads to an augmented neuroinflammatory process that increases the severity of neuropathologic sequelae.


Hyperpermeability of intestinal epithelial monolayers is induced by NO: effect of low extracellular pH.
Nitric oxide (NO.) increases the permeability of Caco-2BBe enterocytic monolayers. Many of the toxic effects of NO. are thought to be mediated by the peroxynitrite anion (ONOO-), which, under mildly acidic conditions, can rearrange to yield an intermediate with reactivity similar to toxic OH.. Accordingly, we assessed the permeability of Caco-2BBe cells grown on permeable supports for 24 h in media titrated to normal or acidic extracellular pH (pHo) with or without the NO. donors 3-morpholinosydnonimine (SIN-1) or sodium nitroprusside (SNP). Incubation with 2 mM SIN-1 at pHo 6.8 or 0.6 mM SNP at pHo 6.5 increased permeability (apical-to-basolateral flux of fluorescein sulfonic acid), whereas at pHo 7.4 permeability was unaffected by these concentrations of NO. donors. Accumulation of NO2/NO3 in medium (index of NO. release) was not increased by incubation of cells with SIN-1 or SNP under mildly acidic conditions. Under acidic but not control conditions, incubation with SIN-1 caused disruption of perijunctional actin filaments as assessed by fluorescence microscopy. At pHo 6.8 and 6.5 (but not 7.4), SIN-1 significantly decreased intracellular levels of both ATP and glutathione. Incubation with 5 mM deferoxamine or 500 uM ascorbic acid (ONOO- scavengers) abrogated SIN-1-induced hyperpermeability. We conclude that mild acidosis augments NO.-induced intestinal epithelial permeability, possibly by promoting oxidant-mediated cytoskeletal damage and/or ATP depletion.